LOCUS       Exported                3987 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Mammalian expression vector pSVSPORTcHLhTNFE, complete sequence.
ACCESSION   LT727394
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3987)
  AUTHORS   De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman 
            Fonseca M, Vanhoucke M, Beyaert R.
  TITLE     BCCM/LMBP Plasmid collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3987)
  AUTHORS   De Schamphelaire W.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, 
            Technologiepark 927, 9052, BELGIUM
REFERENCE   3  (bases 1 to 3987)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3987
                     /organism="Mammalian expression vector pSVSPORTcHLhTNFE"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 3796."
                     /db_xref="taxon:1945141"
     regulatory      1..282
                     /regulatory_class="promoter"
                     /note="SV40 early promoter"
     promoter        10..339
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      190..325
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      239..314
                     /label=SV40 ori
                     /note="SV40 ori"
     promoter        345..363
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     regulatory      345..361
                     /regulatory_class="promoter"
                     /note="SP6 promoter"
     CDS             396..608
                     /codon_start=1
                     /note="unnamed protein product; chicken HL presequence"
                     /protein_id="SJL88333.1"
                     /translation="MDEERLSDNVRLYKGGSIRQGLRSFAAVYVLLALSFLLLTLLSSV
                     SLARIAALSSKLSTLQSEPKHNFSSR"
     CDS             609..1079
                     /codon_start=1
                     /note="unnamed protein product; human TNF mature sequence"
                     /protein_id="SJL88334.1"
                     /translation="VRSSSRTPSDKPVAHVVANPQAEGQLQWLNRRANALLANGVELRD
                     NQLVVPSEGLYLIYSQVLFKGQGCPSTHVLLTHTISRIAVSYQTKVNLLSAIKSPCQRE
                     TPEGAEAKPWYEPIYLGGVFQLEKGDRLSAEINRPDYLDFAESGQVYFGIIAL"
     misc_feature    1089..1127
                     /label=E-tag
                     /note="E-tag"
     promoter        complement(1305..1323)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      complement(1307..1323)
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     misc_feature    1412..1451
                     /label=intron of SV40 small t-antigen
                     /note="intron of SV40 small t-antigen"
     regulatory      complement(1688..1784)
                     /regulatory_class="terminator"
                     /note="SV40 polyA late"
     regulatory      1688..1784
                     /regulatory_class="terminator"
                     /note="SV40 polyA early"
     polyA_signal    1710..1844
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1897..2299)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     CDS             complement(2830..3690)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin resistance"
                     /protein_id="SJL88335.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3691..3795)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      3886..3943
                     /regulatory_class="promoter"
                     /note="lacUV5 promoter"
     protein_bind    3946..3962
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."